The effect of altering the expression level of the F5H gene was investigated in three wood tissues (normal, opposite and tension wood) in 1-year-old hybrid poplar clone 717 (Populus tremula × Populus alba L.), containing the F5H gene under the control of the C4H promoter. Elevated expression of the F5H gene in poplar has been previously reported to increase the percent syringyl content of lignin. The wild-type and three transgenic lines were inclined 45° for 3 months to induce tension wood formation. Tension and opposite wood from inclined trees, along with normal wood from control trees, were analyzed separately for carbohydrates, lignin, cellulose crystallinity and microfibril angle (MFA). In the wild-type poplar, the lignin in tension wood contained a significantly higher percentage of syringyl than normal wood or opposite wood. However, there was no significant difference in the percent syringyl content of the three wood types within each of the transgenic lines. Increasing the F5H gene expression caused an increase in the percent syringyl content and a slight decrease in the total lignin in normal wood. In tension wood, the addition of a gelatinous layer in the fiber walls resulted in a consistently lower percentage of total lignin in the tissue. Acid-soluble lignin was observed to increase by up to 2.3-fold in the transgenic lines. Compared with normal wood and opposite wood, cell wall crystallinity in tension wood was higher and the MFA was smaller, as expected, with no evidence of an effect from modifying the syringyl monomer ratio. Tension wood in all the lines contained consistently higher total sugar and glucose percentages when compared with normal wood within the respective lines. However, both sugar and glucose percentages were lower in the tension wood of transgenic lines when compared with the tension wood of wild-type trees. Evaluating the response of trees with altered syringyl content to gravity will improve our understanding of the changes in cell wall chemistry and ultrastructural properties of normal, opposite and tension wood in plants.